Peptide processing and targeting in the neuronal secretory pathway

The abdominal ganglion of the marine mollusk Aplysia contains a pair of identified neuronal clusters, the bag cells, which control egg laying by means of a number of unique regulatory mechanisms. Each neuron in the bag cell clusters synthesizes several peptides derived from a single prohormone and packages them into separate vesicles. These vesicles are then differentially localized in specific neuronal processes, thus segregating peptides destined for autocrine and hormonal release sites. Therefore in this system, protein trafficking through the secretory pathway organizes multiple peptide neurochemical messengers to efficiently regulate simple behaviors.     

Computed tomographic evaluation of abdominal fat in minipigs

Computed tomography (CT) exams were conducted to determine the distribution of abdominal fat identified based on the CT number measured in Hounsfield Units (HU) and to measure the volume of the abdominal visceral and subcutaneous fat in minipigs. The relationship between the CT-based fat volumes of several vertebral levels and the entire abdomen and anthropometric data including the sagittal abdominal diameter and waist circumference were evaluated. Moreover, the total fat volumes at the T11, T13, L3, and L5 levels were compared with the total fat volume of the entire abdomen to define the landmark of abdominal fat distribution. Using a single-detector CT, six 6-month-old male minipigs were scanned under general anesthesia. Three radiologists then assessed the HU value of visceral and subcutaneous abdominal fat by drawing the region of interest manually at the T11, T13, L1, L3, and L5 levels. The CT number and abdominal fat determined in this way by the three radiologists was found to be correlated (intra-class coefficient = 0.9). The overall HU ranges for the visceral and subcutaneous fat depots were -147.47 to -83.46 and -131.62 to -90.97, respectively. The total fat volume of the entire abdomen was highly correlated with the volume of abdominal fat at the T13 level (r = 0.97, p < 0.0001). These findings demonstrate that the volume of abdominal adipose tissue measured at the T13 level using CT is a strong and reliable predictor of total abdominal adipose volume.     

Analysis of methanogenic archaeal communities of rumen fluid and rumen particles from Korean black goats

Molecular diversity of methanogens in the rumen of Korean black goats was investigated with 16S rRNA gene clone libraries using methanogen‐specific primers. The libraries were composed of rumen fluid‐associated methanogens (FAM) and rumen particle‐associated methanogens (PAM) from rumen‐fistulated Korean black goats. Among the 141 clones of the FAM library, the sequences were mostly related to two phyla, the Methanobacteriaceae family (77.3%) and the Thermoplasmatales family (22.7%); and among the 68 clones of the PAM library, sequences were also mainly clustered in the two phyla, the Thermoplasmatales family (63.24%) and the Methanobacteriaceae family (35.29%). Most of the sequenced clones in the two libraries were closely related to uncultured methanogenic archaeon. Quantitative real‐time PCR revealed that PAM (8.97 log 10) had significantly higher (P < 0.01) density of methanogens by the methanogenic 16S rRNA gene copies than FAM (7.57 log 10). The two clone libraries also showed difference in Shannon index (FAM library 1.70 and PAM library 1.59) and Chao 1 estimator (FAM library 18 and PAM library 17 operational taxonomic units). Apparent differences found in the microbial community from the two 16S rRNA gene libraries could be a result of such factors as the chemical and physical nature of the target material surface, types or component of diets, the interaction between the methanogens and other microbes, and age of the experimental goats.     

Using metabolic profiling to assess plant-pathogen interactions: an example using rice (<Emphasis Type="Italic">Oryza sativa)</Emphasis> and the blast pathogen <Emphasis Type="Italic">Magnaporthe grisea</Emphasis>

A metabolomics based approach has been used to study the infection of the Hwacheong rice cultivar (Oryza sativa L. cv. Hwacheong) with compatible (KJ201) and incompatible (KJ401) strains of the rice blast fungal pathogen Magnaporthe grisea. The metabolic response of the rice plants to each strain was assessed 0, 6, 12, 24, 36, and 48 h post inoculation. Nuclear Magnetic Resonance (NMR) spectroscopy and Gas and Liquid Chromatography Tandem Mass spectrometry (GC/LC-MS/MS) were used to study both aqueous and organic phase metabolites, collectively resulting in the identification of 93 compounds. Clear metabolic profiles were observed at each time point but there were no significant differences in the metabolic response elicited by each pathogen strain until 24 h post inoculation. The largest change was found to be in alanine, which was ~30% (±9%) higher in the leaves from the compatible, compared to the resistant, plants. Together with several other metabolites (malate, glutamine, proline, cinnamate and an unknown sugar) alanine exhibited a good correlation between time of fungal penetration into the leaf and the divergence of metabolite profiles in each interaction. The results indicate both that a wide range of metabolites can be identified in rice leaves and that metabolomics has potential for the study of biochemical changes in plant-pathogen interactions.     

Cardiomyocyte calcium cycling in a naturally occurring German shepherd dog model of inherited ventricular arrhythmia and sudden cardiac death

ObjectiveTo further characterize arrhythmic mechanisms in German shepherd dogs (GSDs) affected with inherited ventricular arrhythmias by evaluating intracellular calcium cycling and expression of calcium handling genes.AnimalsTwenty five GSDs, 9 backcross dogs, and 6 normal mongrel dogs (controls) were studied. The GSDs and backcross dogs were from a research colony of inherited ventricular arrhythmias. The control research dogs were purchased.MethodsAction potentials (APs) and pseudo-electrocardiograms (ECG) were recorded from left ventricular (LV) wedge preparations of GSDs and normal dogs. Midmyocardial (Mid) LV cells from GSDs and normal mongrels were isolated by enzymatic digestion. Cells were either field stimulated or voltage clamped and calcium transients were measured by confocal microscopy using the indicator Fluo-3AM. Expression of calcium handling genes was measured by quantitative RT-PCR.ResultsMean calcium transient decay (tau) was not different between affected GSDs and control dogs, but striking cell-to-cell variability for tau was observed within affected GSDs and between affected GSDs and controls (P < 0.0001 each); within-dog variability accounted for 75% of total variability. Calcium sparks and afterdepolarizations occurred in GSD but not control cells. ATP2A2/SERCA2a expression was significantly reduced (P = 0.0063) in affected GSDs and inversely correlated (P = 0.0006) with severity of ventricular arrhythmias.ConclusionsGerman shepherd dogs with inherited ventricular arrhythmias have electrophysiologic abnormalities in calcium cycling associated with reduced ATP2A2/SERCA2a expression. These animals provide a unique opportunity to study calcium remodeling at the genetic and molecular level in familial ventricular arrhythmias.     

Infectious aetiologies of mastitis on Korean dairy farms during 2008

The objective of this study was to determine the frequency of different mastitis pathogens in 1255 milk samples collected from 368 lactating cows on 24 dairy farms in Korea during 2008. The proportion of cows and quarter milk samples having SCC ? 200,000 cells/ml, an indicator of udder infection, was 54.3% (200/368 cows) and 35.5% (446/1255 samples), respectively. Of the 446 milk samples subjected to bacteriological examination, 16.5% (74) showed no bacterial growth and 3.5% (16) were contaminated. In total, 356 of 1255 (28.3%) samples were bacteriologically positive, from which 415 bacteria were isolated. The most frequently isolated pathogen was coagulase-negative staphylococci (40.7%), followed by Gram-negative bacteria (19.5%) other than Escherichia coli, Staphylococcus aureus (12.2%), Streptococcus uberis (5.3%), Enterococcus spp. (4.8%), E. coli (4.5%), and environmental streptococci (3.1%) other than S. uberis. This study demonstrates that environmental pathogens were the vast majority of bacteria isolated from mastitic bovine milk samples in Korea.     

Genotypic prevalence of the adhesin involved in diffuse adherence in Escherichia coli isolates in pre-weaned pigs with diarrhoea in Korea

A total of 1002 Escherichia coli strains isolated from pre-weaned pigs with diarrhoea on 1114 swine farms were screened for the presence of the adhesin involved in diffuse adherence (AIDA) gene by polymerase chain reaction (PCR). Escherichia coli isolates that carried AIDA genes were also tested by PCR for the detection of five fimbriae (F4, F5, F6, F18 and F41), heat-stable (STa, STb) and heat-labile (LT) enterotoxin, enteroaggregative E. coli heat-stable enterotoxin 1 (EAST1), and Shiga toxin 2 oedema disease (Stx2e) genes. Twenty-three (2.3%) of the 1002 E. coli isolates carried the gene for AIDA. Among 23 isolates shown to carry genes for AIDA, three carried the AIDA gene as the only shown virulence factor. Other isolates carried other virulence factor genes in addition to AIDA. Four isolates carried genes for at least one of the fimbrial adhesins and enterotoxins. Sixteen isolates carried genes for enterotoxins only. The AIDA may represent an additional virulence determinant in pre-weaned pigs with diarrhoea.     

Influence of Oil Seed Rape and Faba Bean as Green Manure on Nitrogen Utilization in a Spring Cereal Rotation

A lysimeter study was conducted to investigate how nitrogen uptake of spring wheat and oats and N leaching losses are differing when either oilseed rape or faba bean are grown as green manure. The experiment was conducted during 1982–1988, and fertilizer applied to the cereals was either farmyard manure, FYM, (6 and 12 g N/m²/year) or mineral fertilizer, MiF, at a rate of 6 g N/m². On average of the 7 year period faba bean increased N uptake of the cereals fertilized with FYM at harvest by 3–4 g/m². Differences between the two application rates were negligible. N-leaching losses increased twofold when faba bean was grown instead of oilseed rape. Growing faba bean after cereals supplied with MiF, too, increased N uptake of the cereals and significantly increased leaching losses.     

Effects of dietary fiber of young adult genetically lean, obese and contemporary pigs: rate of passage, digestibility and microbiological data

Twenty-one genetically lean, obese or contemporary slaughter weight castrated male pigs (6 mo old; seven of each genotype) were assigned to individual tether stalls and fed either a control diet (low fiber) or a diet containing 80% alfalfa meal (high fiber) at 1.50% of initial body weight for 71 d (1.75% for d 1 to 4). Apparent dry matter digestibility of the diets was estimated by determining acid insoluble ash in fecal samples. Fecal cellulolytic bacteria and total viable bacteria were enumerated at d 0, 14, 35, 49 and 70. Fecal inocula were used to determine 48-h in vitro digestibility of alfalfa meal fractions on the same days. Digesta rate of passage was determined by feeding a pulse dose of chromium-mordanted alfalfa fiber to the pigs fed the high-fiber diet. In vivo digestibility of both diets was less for the obese pigs than for the lean or contemporary genotypes. In vitro digestibility of alfalfa fiber fractions was not different between the genotypes fed either diet. When the high-fiber diet was fed, in vitro digestibility increased for all genotypes from d 0 to d 14, but not thereafter. The numbers of cellulolytic bacteria for all three genotypes were greater when pigs were fed the high-fiber diet (23.0 X 10(8), 51.6 X 10(8), 37.2 X 10(8) per gram fecal dry weight; obese, lean and contemporary, respectively) compared to the low-fiber diet (3.0 X 10(8), 3.2 X 10(8), 3.4 X 10(8), respectively).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of dietary fiber on young adult genetically lean, obese and contemporary pigs: body weight, carcass measurements, organ weights and digesta content

Twenty-one genetically lean, obese or contemporary barrows (6 mo old; seven of each genotype) were assigned to individual tether stalls and fed a control diet (low-fiber) or a diet containing 80% alfalfa meal (high-fiber) at 1.50% of initial body weight for 71 d (1.75% for d 1 to 4). Backfat thickness was recorded ultrasonically at 2-wk intervals, and body weight was recorded at the beginning and end of the 10-wk experiment. Pigs were slaughtered after a 24-h fast, and carcass weight, length and backfat thickness and cross-sectional area of the longissimus muscle were measured. Weights of cecum, heart, liver and kidney and of full and empty stomach and colon and empty small intestine were recorded. Volume and weight of colon and cecum contents were determined. Restriction of digestible energy reduced weight gain to zero or below in pigs fed alfalfa meal compared with 220 g daily in pigs fed the low-fiber diet. Restriction of energy reduced backfat in all three genotypes. Liver, kidney and empty segments of the gastrointestinal tract as a percentage of body weight were increased by high fiber. Obese pigs had smaller longissimus muscle area, more backfat and smaller liver, heart, empty stomach and colon than lean or contemporary pigs, but there were no diet X genotype interactions for any of these traits. Obese pigs consistently had smaller digesta volumes and dry matter weights than the other genotypes. The increased relative organ weights and the associated disproportionate contribution of these organs to body energy expenditure have important implications for effects on basal metabolic rate.(ABSTRACT TRUNCATED AT 250 WORDS)